Tsim cov Fluorescence Qis thiab Bright-Field Microscopes
Kauj ruam 1: Txheej txheem cej luam ntawm Fluorescence Microscopy
Txhawm rau nkag siab txog cov ntsiab lus tseem ceeb ntawm fluorescence microscopy, xav txog lub hav zoov ntom ntom thaum hmo ntuj nrog ntoo, tsiaj txhu, tsob ntoo, thiab lwm yam hav zoov nyob hauv nws. Yog tias koj ci lub teeb nyem rau hauv hav zoov, koj yuav pom tag nrho cov qauv no thiab nws nyuaj rau xav txog tej yam tsiaj lossis nroj tsuag. Piv txwv tias koj tsuas yog xav pom blueberry shrubs hauv hav zoov. Yuav kom ua tiav qhov no, koj yuav tsum tau cob qhia fireflies kom tsuas yog nyiam rau blueberry bushes, yog li ntawd thaum koj saib tom hav zoov, tsuas yog blueberry bushes yuav teeb. Koj tuaj yeem hais tias koj siv fireflies los kos blueberry bushes, kom koj tuaj yeem pom cov qauv blueberry hauv hav zoov.
Hauv qhov piv txwv no, hav zoov sawv cev rau tag nrho cov qauv, blueberry shrubs sawv cev rau cov qauv uas koj xav pom (xws li cov hlwb tshwj xeeb lossis cov kab mob subcellular), thiab cov kab hluav taws xob yog cov fluorescent compounds. Qhov xwm txheej ntawm kev teeb pom kev ntawm tus neeg tua lub teeb nyem ib leeg yam tsis muaj hluav taws zoo ib yam li lub teeb pom kev zoo.
Cov kauj ruam tom ntej yog kom nkag siab txog cov haujlwm tseem ceeb ntawm cov tshuaj fluorescent (tseem hu ua fluorophores). Fluorescent pawg yog cov khoom me me (ntawm nanoscale) tsim los txuas cov qauv tshwj xeeb hauv cov qauv. Lawv nqus lub teeb ntawm qhov nqaim ntawm wavelengths thiab rov emit lub teeb ntawm lwm lub wavelength. Piv txwv li, fluorophore tuaj yeem nqus tau lub teeb xiav (xws li cov fluorophore zoo siab los ntawm lub teeb xiav) thiab tom qab ntawd rov tawm lub teeb ntsuab. Feem ntau, qhov no yog sau los ntawm excitation thiab emission spectra (raws li qhia hauv daim duab saum toj no). Cov kab kos no qhia txog lub wavelengths ntawm lub teeb absorbed los ntawm fluorophores thiab lub wavelengths ntawm lub teeb emitted los ntawm fluorophores.
Lub tshuab microscope tsim zoo ib yam li lub tshuab tsom iav tsis tu ncua, nrog ob qhov sib txawv tseem ceeb. Ua ntej, lub teeb illuminating tus qauv yuav tsum yog ntawm lub wavelength uas excites pawg fluorescent (piv txwv li saum toj no, lub teeb yog xiav). Qhov thib ob, lub tshuab tsom iav tsuas yog xav tau los sau cov teeb pom kev zoo (lub teeb ntsuab) thaum thaiv lub teeb xiav. Qhov no yog vim lub teeb xiav nyob txhua qhov chaw, tab sis lub teeb ntsuab tsuas yog los ntawm cov qauv tshwj xeeb hauv cov qauv. Txhawm rau thaiv lub teeb xiav, microscopes feem ntau muaj qee yam hu ua lub lim dej ntev uas tso cai rau lub teeb ntsuab hla dhau yam tsis muaj lub teeb xiav. Txhua lub lim lim qis muaj qhov txiav tawm wavelength. Yog hais tias lub wavelength ntawm lub teeb ntau dua li qhov kev txiav tawm wavelength, nws tuaj yeem dhau los ntawm lub lim. Yog li ntawd, lub npe yog 'Long Distance Pass'. Luv wavelengths raug thaiv.
Kauj Ruam 2: Siv lub tshuab ntsuas qhov muag pom
Qhov no yog ib kauj ruam ntxiv hauv kev tsim cov tshuab microscopes raws li cov hauv paus ntsiab lus. Tsis tas yuav tsim lub fluorescence microscope, yog li yog tias koj tsis xav delve tob rau hauv optics, koj tuaj yeem hla nws.
Ob qho chaw ci ntsa iab thiab fluorescence microscopes tuaj yeem ua qauv siv cov khoom siv hluav taws xob optics. Lub hauv paus ntsiab lus ntawm ray optics yog tias tus cwj pwm ntawm lub teeb zoo ib yam li lub teeb propagating deb ntawm lub teeb. Thaum koj saib ib ncig ntawm chav tsev, koj yuav pom lub hnub ci sab nraum lub qhov rais lossis lub teeb coj los ntawm lub teeb. Tom qab ntawd lub teeb yog absorbed los yog reflected los ntawm cov khoom nyob rau hauv lub chav tsev. Qee lub teeb pom kev zoo yuav coj nws mus rau koj lub qhov muag. Yog hais tias ib yam khoom yog illuminated, koj muaj peev xwm xav tias txhua qhov taw tes ntawm cov khoom emits lub teeb nyob rau hauv tag nrho cov lus qhia (raws li qhia nyob rau hauv daim duab saum toj no). Lub lens, zoo li lub lens hauv peb lub qhov muag, tsom lub teeb rau ntawm qhov taw tes kom peb pom cov khoom. Yog tsis muaj lub lens, lub teeb txuas ntxiv mus rau sab nraud thiab tsis tsim cov duab.
